Table 1.
Sulfolobus strains and vectors used in this study
| Strain or vector | Relevant characteristics | Source or reference |
|---|---|---|
| Sulfolobus strains | ||
| S. islandicus REY15A | Wild type | 7 |
| S. islandicus E233S | REY15A ΔpyrEF ΔlacS | 8 |
| S. solfataricus P2 | Wild type | 20 |
| Vectors | ||
| pZC1 | Sulfolobus-E. coli shuttle vector derived from pHZ2 | 18 |
| pDL1 | Derived from pZC1, lacking PvuII, NdeI, and ZraI sites | 10 |
| pRp | Sulfolobus reporter plasmid constructed with pZC1, containing an arabinose-inducible promoter (−55, +6) and lacS encoding a β-glycosidase as the reporter gene | 18 |
| pEXA | Sulfolobus expression vector derived from pZC1, containing a core araS promoter (ParaS1, −109, +6), an MCS, and a hexahistidine-coding sequence at each terminus, as well as a transcriptional terminator (T6 of SSV1) | 10 |
| pEXA-N-lacS | S. solfataricus lacS was inserted into pEXA to yield a fusion gene coding for N-terminal His tag-LacS fusion protein | This study |
| pEXA-C-lacS | S. solfataricus lacS was inserted into pEXA to yield a fusion gene coding for C-terminal His tag-LacS fusion protein | This study |
| pEXA-N/C-lacS | S. solfataricus lacS was inserted into pEXA to yield a fusion gene coding for a LacS fusion protein fused to a His tag at both ends | This study |
| pSeSD | The synthetic araS-SD promoter was used to substitute for ParaS2 in pEXA | This study |
| pSeSD-estA | An estA gene (SiRe_0290; named pSe-estA in reference 17) was inserted into pSeSD | 17 |
| pSeSD-N-lacS | S. solfataricus lacS was inserted into pSeSD to yield a fusion gene coding for an N-terminal His tag-LacS fusion protein, as for pEXA-N-lacS | This study |
| pSeSD-C-lacS | S. solfataricus lacS was inserted into pSeSD to yield a fusion gene coding for a C-terminal His tag-LacS fusion protein | This study |
| pSeSD-N/C-lacS | S. solfataricus lacS was inserted into pSeSD to yield a fusion gene coding for a LacS fusion protein fused to a His tag at both ends | This study |
| pSeTL1 | ParaS2 of pEXA was replaced with an extended ParaS (−55, +39), the araS promoter plus the coding sequence of the first 11 amino acids of the arabinose-binding protein | This study |
| pSeTL1-estA | S. islandicus estA was inserted into pSeTL1 | This study |
| pSeTL2 | ParaS2 of pEXA was replaced with an extended ParaS (−55, +24), the araS promoter plus the coding sequence of first 6 amino acids of the arabinose-binding protein | This study |
| pSeTL2-estA | S. islandicus estA was inserted into pSeTL2 | This study |
| p10b | The sso-alba gene promoter (−192, +8) was used to substitute for ParaS2 in pEXA | |
| p10b-C-lacS | S. solfataricus lacS was inserted into p10b | This study |
| p7d | The sso7d gene promoter (−231, +7) was used to substitute for ParaS2 in pEXA | This study |
| p7d-C-lacS | S. solfataricus lacS was inserted into p7d | This study |